Multiplex PCR for the identification of Arcobacter and differentiation of Arcobacter butzleri from other arcobacters.
A multiplex polymerase chain reaction (PCR) assay to identify Arcobacter isolates and to distinguish A. butzleri from other arcobacters is described. The test uses two primer sets. Set I targets a section of the 16S rRNA genes of Arcobacter spp. Set II amplifies a portion of the 23S rRNA genes unique to A. butzleri. Specificity of the primer sets was evaluated using ATCC reference strains of A. butzleri, A. cryaerophilus, A. skirrowii, Bacteroides spp., Campylobacter spp., Helicobacter spp. and Wolinella succinogenes. Upon PCR amplification, all of the Arcobacter isolates yielded a 1223 bp product, whereas A. butzleri ATCC 49616 exhibited both a 1223 bp and a 686 bp product. No PCR product was observed for other closely related ATCC strains (n = 37). We next analyzed by multiplex PCR field strains of Arcobacter spp. (n = 108) which had been previously characterized to the species level by either DNA-DNA hybridization, dot blot hybridization, ribotyping or by serology. The 1223 bp multiplex PCR product identified all of the isolates as Arcobacter. The presence of both the 1223 and 686 bp amplicons identified 66 strains as A. butzleri. Speciation by multiplex PCR agreed with results obtained by the other methods. The multiplex PCR assay is specific, rapid and easy to interpret and, thus, will aid in elucidating the prevalence, epidemiology and zoonotic potential of Arcobacter.
Harmon
K M
KM
Enteric Diseases and Food Safety Research Unit, National Animal Disease Center, USDA, Ames, IA 50010, USA.
Wesley
I V
IV
D000818:Animals; D001439:Bacteroides / Q000145:classification; D002167:Campylobacter / Q000145:classification; D002645:Chickens / Q000382:microbiology; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016905:Gram-Negative Bacterial Infections / Q000145:classification / Q000175:diagnosis / Q000662:veterinary*; D016998:Helicobacter / Q000145:classification; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry / Q000382:microbiology*; D011201:Poultry Diseases; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012703:Serotyping; D014422:Turkeys / Q000382:microbiology; D016968:Wolinella / Q000145:classification
D012336:RNA, Ribosomal, 16S